Immunological and Symptomatic Effects of Oral Intake of Transgenic Rice Containing 7 Linked Major T-Cell Epitopes from Japanese Cedar Pollen Allergens.

BACKGROUND: A rice-based peptide vaccine containing 7 linked human predominant T-cell epitopes (7Crp) derived from Japanese cedar (JC) pollen allergens, Cry j 1 and Cry j 2, was developed. Here, we examined the efficacy and safety of this transgenic rice in JC pollinosis patients. METHODS: Transgenic rice (5, 20, and 80 g) was administered orally. We measured the T-cell proliferative activity against 7Crp, Cry j 1, and Cry j 2; the cytokine expression levels; and specific IgE and IgG4 production levels. In addition, the symptom and medication scores were monitored during the pollen season, and quality of life (QOL) was evaluated. RESULTS: T-cell proliferative activities to Cry j 1, Cry j 2, and 7Crp were significantly depressed in a dose-dependent manner. Oral intake of 80 g transgenic rice for 20 weeks resulted in significant suppression of allergen-specific T-cell proliferation with downregulation of IL-13 and upregulation of IL-10 levels but no changes to specific IgE and IgG4 levels. The QOL symptom scores for allergic rhinitis were not significantly improved. CONCLUSIONS: Allergen-specific T-cell responses were significantly reduced by oral intake of transgenic rice in a dose-dependent manner. However, neither medication score nor QOL symptom scores could be improved during the JC pollen season with oral intake of transgenic rice for 20 weeks.

Long-term oral administration of transgenic rice containing cedar pollen T-cell epitopes potentially improves medication- and allergy-related quality-of-life scores.

Background: We previously developed a transgenic rice that contains seven linked human predominant T-cell epitopes (7Crp) derived from Japanese cedar (JC) pollen allergens Cry j 1 and Cry j 2. Oral administration of 80 g of transgenic rice for 20 weeks suppressed allergen-specific T-cell proliferation in participants with JC pollinosis, but their clinical symptoms did not improve. Objective: We examined the clinical efficacy of low-dose (5 g and 20 g) intake of the transgenic rice administered for two successive seasons. Methods: In this randomized, double-blind, placebo controlled study, transgenic rice seeds (5 g or 20 g) were orally administered to the participants for 24 weeks in each of two successive JC pollen seasons. We analyzed T-cell proliferation and cytokine expression, and monitored symptom and medication scores during the pollen season. Quality of life (QOL) was evaluated by using the Japanese Allergic Rhinitis Quality of Life Standard Questionnaire (JRQLQ). Results: Specific T-cell proliferation after stimulation with 7Crp, Cry j 1, and Cry j 2 was significantly suppressed in the second JC pollen season. No significant differences were found among the three groups (5 g, 20 g, and placebo) with regard to clinical symptoms or medication scores in the first season. However, the medication scores and face scale for overall condition of JRQLQ improved in the 5-g transgenic rice group in the second season, although careful re-examination with a large sample size is necessary to confirm the results. Conclusion: Low-dose oral administration of transgenic rice that contains 7Crp significantly reduced allergen-specific T-cell responses and improved medication scores during the second season of administration. Thus, oral intake of the transgenic rice has the potential to induce immune tolerance to JC pollen allergens when administered for at least two successive seasons.

T-cell activation by transgenic rice seeds expressing the genetically modified Japanese cedar pollen allergens.

Transgenic rice seeds that contain genetically modified Cry j 1 and Cry j 2, the two major allergens of Cryptomeria japonica (Japanese cedar; JC), have been developed as immunotherapeutic candidates for JC pollinosis. Because the transgenic rice (TG-rice) seeds express allergens containing whole amino acid sequences of Cry j 1 and Cry j 2 in the endosperm tissue (edible part of rice grain), they can potentially target all Cry j 1- and Cry j 2-specific T-cells. However, it was unknown whether antigenicity of Cry j 1 and Cry j 2 could be completely preserved in TG-rice seeds. We verified the antigenicity of TG-rice seeds to T-cells through the analysis of the proliferative responses of T-cells in Cry j 1- or Cry j 2-immunized mice or T-cell lines to TG-rice seed extract. First, four mouse strains were immunized with Cry j 1 or Cry j 2. T-cells in the immunized mice proliferated on treatment with TG-rice seed extract, but not non-transgenic wild-type rice (WT-rice) seed extract. Furthermore, T-cell lines were established from the spleen cells of the immunized mice. Each T-cell line resulted in a proliferative response to TG-rice seed extract, but not to WT-rice seed extract, suggesting that TG-rice seeds certainly express T-cell epitopes corresponding to T-cell lines. Considering the modified amino acid sequences of Cry j 1 and Cry j 2 in TG-rice seeds, the expression of specific T-cell epitopes suggested that TG-rice seeds express all possible T-cell epitope repertoires of Cry j 1 and Cry j 2.

Impact of residual frontal recess cells on frontal sinusitis after endoscopic sinus surgery.

PURPOSE: Endoscopic sinus surgery (ESS) is a well-established treatment for chronic rhinosinusitis (CRS). However, ESS for frontal sinusitis remains complicated and challenging. The aim of this study was to identify the relationship between residual frontal recess cells and primary ESS failure in the frontal sinus. METHODS: We prospectively collected information on 214 sides of 129 patients with CRS who underwent standard ESS from June 2010 to May 2011. To identify risk factors, we retrospectively analyzed clinical data and computed tomography (CT) images before and 3 months after surgery. RESULTS: The posterior side of the frontal recess cells remained relatively common: suprabullar cells (SBCs) were found in 12.2% (16 sides), suprabullar frontal cells (SBFCs) in 20.3% (12 sides), and supraorbital ethmoid cells in 23.7% (14 sides). In contrast, the anterior side of the frontal recess cells, agger nasi cells, supra agger cells, and supra agger frontal cells remained at < 10.0%. Frontal septal cells persisted in 25.0% (5 sides). The presence of residual frontal recess cells was an independent risk factor for postoperative frontal sinus opacification as were well-recognized risk factors such as nasal polyps, the peripheral eosinophil count, and the CT score. Among residual frontal recess cells, SBCs and SBFCs were independent risk factors for opacification. CONCLUSIONS: Residual frontal recess cells, especially SBCs and SBFCs, were independent risk factors for postoperative opacification of the frontal sinus. Complete surgical excision of frontal recess cells may improve surgical outcomes.

Nasal silicone splints and quilting sutures using a septum stitch device following septoplasty: a prospective comparative study.

PURPOSE: Although nasal splints and trans-septal sutures are preferred for septal closure following septoplasty, quilting sutures, also known as continuous mattress sutures, are technically challenging and thus time consuming. Recently, the utility of a novel stitch device was demonstrated for quilting sutures; however, whether it contributes to the technical solution and time management of septal sutures remains unclear. This study aims to quantify the time and cost of septal closure following septoplasty by comparing nasal splints with quilting sutures using the septum stitch device. METHODS: We prospectively examined 23 patients who underwent septoplasty and assigned them to the following two groups: group 1, underwent the quilting suture for septal closure following septoplasty; group 2 underwent the nasal silicone splint method. We recorded the total operation time and surgical time for septal closure. Furthermore, we evaluated postoperative symptoms with Visual Analogue Scale (VAS) scores and compared the complication rate. RESULTS: We observed no significant difference in the average of total operating time (P = 0.641) and the surgical time for septal closure (P = 0.886). Among postoperative symptoms, only nasal bleeding was significantly worse in group 1 on postoperative day 1 (P = 0.023). In addition, the occurrence of postoperative complications was insignificant between both groups. CONCLUSIONS: The septum stitch facilitated safe and easy quilting suturing without significant increase of operation time.

Endoscopic modified medial maxillectomy for odontogenic cysts and tumours.

BACKGROUND: Odontogenic maxillary cysts and tumours originate from the tooth root and have traditionally been treated through an intraoral approach. Here, we report the efficacy and utility of endoscopic modified medial maxillectomy (EMMM) for the treatment of odontogenic maxillary cysts and a tumour. METHODOLOGY: We undertook EMMM under general anaesthesia in six patients: four had radicular cysts, one had a dentigerous cyst, and one had a keratocystic odontogenic tumour. RESULTS: The cysts and tumours were completely excised and the inferior turbinate and nasolacrimal duct were preserved in all patients. There were no peri- or postoperative complications, and no incidences of recurrence. CONCLUSION: Endoscopic modified medial maxillectomy appears to be an effective and safe technique for treating odontogenic cysts and tumours.

The expression of protease-activated receptors in chronic rhinosinusitis.

BACKGROUND: A recent study suggested that protease-activated receptors (PARs) are involved in allergic respiratory diseases, such as asthma. Chronic rhinosinusitis (CRS) is one of the most common chronic airway diseases, but little is understood about its pathogenesis. The purpose of this study was to compare the expression and distribution of PARs in biopsy specimens obtained from CRS and control patients. METHODS: Biopsy specimens were obtained from 7 pituitary tumor patients as controls, 8 CRS patients with aspirin-tolerant asthma (ATA), 7 CRS patients with aspirin-induced asthma (AIA), and 7 CRS patients without asthma (CRS). Sections were stained for PAR-1, PAR-2, PAR-3 and PAR-4 using specific polyclonal antibodies. Staining was scored semiquantitatively for both intensity and distribution. To confirm the presence of PARs on inflammatory cells, double staining with eosinophil cationic protein (EG2) and elastase was also performed. RESULTS: Both the epithelium and the infiltrating inflammatory cells in the CRS with asthma groups showed significant upregulation of the expression of PAR-2 and PAR-3 compared with the CRS without asthma group and the control group. In the patients with CRS complicated by asthma, eosinophils were increased among PAR-2- and PAR-3-positive cells. In the patients with CRS not complicated by asthma, neutrophils were increased among PAR-2-positive cells. CONCLUSIONS: Differences in the expression of PAR-2 and PAR-3 on epithelial cells, eosinophils and neutrophils may be involved in the pathogenesis of CRS. CRS may be able to be treated by targeting PAR-2 and PAR-3.

Increased CXCL10 expression in nasal fibroblasts from patients with refractory chronic rhinosinusitis and asthma.

BACKGROUND: Chronic rhinosinusitis (CRS) is characterized by local inflammation of the sinonasal tissues. CRS patients with nasal polyps and asthma often develop acute exacerbation of sinonasal symptoms after upper respiratory tract infections. However, the influence of concomitant asthma on the nasal immune response to viral infection remains unclear. METHODS: Specimens of nasal polyp and mucosal tissues were obtained from 3 groups of CRS patients (n = 14 per group): 1) patients without asthma (CRS group), 2) patients with aspirin-tolerant asthma (ATA group), and 3) patients with aspirin-intolerant asthma (AIA group). Nasal fibroblasts isolated from the specimens were stimulated with poly I:C. CXCL10 expression was analyzed by the quantitative real-time polymerase chain reaction and enzyme-linked immunoadsorbent assay. Biopsy specimens from CRS patients without asthma were subjected to immunohistochemistry for detection of T-bet and GATA-3 expression in CD3+ T cells by double labeling. RESULTS: Nasal fibroblasts from the ATA and AIA groups showed significantly enhanced expression of CXCL10 mRNA and protein after poly I:C stimulation compared with cells from the CRS group and the control group (normal nasal mucosa). In addition to T helper (Th)2 cells, there was more abundant infiltration of Th1 cells into tissues from the AIA and ATA groups. CONCLUSIONS: Our findings suggest that CRS associated with asthma may become intractable through the over-production of CXCL10 in response to viral infection.

Elevated levels of interleukin-33 in the nasal secretions of patients with allergic rhinitis.

BACKGROUND: Interleukin (IL)-33, which is a member of the IL-1 family of cytokines, is now recognized as an important contributor to Th2-type immune responses. We examined whether the levels of IL-33 in sera and nasal secretions are upregulated in allergic rhinitis (AR) patients, and we tested for correlations between the IL-33 level and the parameters of atopy and the nasal symptom score. METHODS: The study included 24 Japanese cedar pollinosis patients (12 male and 12 female patients with a mean age of 47.7 years) with a history of moderate-to-severe AR, 14 house-dust-mite-sensitized patients with AR (9 male and 5 female patients with a mean age of 42 years) and 8 normal controls. We used Japan Rhinoconjunctivitis Quality-of-Life Questionnaire sheets to evaluate the nasal symptoms. We collected sera and nasal secretions to examine the level of IL-33 protein by ELISA. RESULTS: IL-33 protein was not detected in the serum of any of the subjects. However, the IL-33 level in nasal secretions was significantly elevated in patients with Japanese cedar pollinosis at peak season and in patients with perennial AR compared to Japanese cedar pollinosis patients at preseason and the normal controls. Furthermore, IL-33 in nasal secretions correlated significantly with the total nasal symptom score. CONCLUSIONS: Our results suggest that IL-33 in nasal secretions may be related to exacerbation of AR, including that of Japanese cedar pollinosis cases.

[Evaluation of clinical features of antrochoanal polyps].

Current knowledge on the prevalence and clinical features of antrochoanal polyps (ACPs), benign lesions arising in the maxillary sinus and extending into the choana, is very limited in Japan. We prospectively evaluated prevalence and clinical features in 15 subjects with ACPs from among 728 undergoing endoscopic endonasal sinus surgery between April 2007 and March 2008, and prospectively enrolled in this study. The 15 subjects, who accounted for 2.1% of the total, had nasal obstruction, rhinorrhea, and postnasal drip. Symptoms significantly reduced postoperatively. Maxillary-sinus-origin ACP distribution was 40% from the maxillary sinus floor to the posterior wall, 26.7% from maxillary sinus floor, and 20% from the maxillary sinus floor to the internal wall. Postoperative recurrence was 13.3%. Endoscopic endonasal sinus surgery for ACPs was most effective for polyp is originating in the maxillary sinus determined carefully and excised as completely as possible, followed by appropriate postoperative treatment.

[Endoscopic sinus surgery complications a prospective multicenter study].

OBJECTIVE: Our objective was to determine the rate of complications in endoscopic sinus surgery (ESS) and associated risk factors. METHODS: We prospectively studied 1,382 subjects undergoing ESS for rhinosinusitis and cystic sinus disease at 16 hospitals during 2007 and 2008. Surgeons provided information on peri-and postoperative complication occurrence. RESULT: Results of complications were seen in 80 subjects (5.8%), the most frequent was perioperative lamina papyracea injury. Analysis showed the complication rate to be linked to gender, and anesthesia type, but not the grade of surgeon. CONCLUSIONS: While care should be taken to avoid them, complications should be identified and treated in a timely and accurate manner.

Management of Intractable Petrous Bone Cholesteatoma With a Combined Translabyrinthine-Transsphenoidal Approach.

OBJECTIVE: To evaluate whether a combined translabyrinthine-transsphenoidal approach can be used to achieve adequate surgical resection of an extensive petrous bone cholesteatoma and create a debris drainage route for the residual cholesteatoma that is maintained long-term. PATIENT: A 71-year-old man with residual petrous temporal bone cholesteatoma that had spread extensively to the internal carotid artery and posterior cranial fossa. INTERVENTION: Surgical resection of the cholesteatoma via a translabyrinthine approach and creation of a debris drainage route into the nasopharynx via a transsphenoidal approach. MAIN OUTCOME MEASURES: Control of unresectable petrous temporal bone cholesteatoma and occurrence of cholesteatoma- or surgery-related complications. RESULTS: Although complete removal of the cholesteatoma was attempted via a translabyrinthine approach, this was not possible because the epithelium of the cholesteatoma was strongly attached to the internal carotid artery and posterior cranial fossa. A debris drainage route leading to the nasopharynx was created by drilling the clivus on the side of the lesion via a transsphenoidal approach. The patient has had no complications since surgery. The drainage route remains open, and the cholesteatoma has been controlled for 57 months. CONCLUSIONS: A markedly advanced petrous temporal bone cholesteatoma can be managed safety and reliably by combining a translabyrinthine approach with a transsphenoidal approach. Creation of a debris drainage route into the nasopharynx can prevent isolation of the cholesteatoma and mastoid cavity problems after surgery. This one-stage surgery may be a suitable method for keeping cholesteatoma under control in patients with unresectable petrous bone cholesteatoma.

Discussion on effect of material on UV reflection and its disinfection with focus on Japanese Stucco for interior wall.

Research has previously shown that ultraviolet light C (UV-C) can inactivate unexpected infection. However, this type of potential disinfection is dramatically reduced for the shadow area such as under desk or medical equipment. Because the UV-C reflectance ratio is low on the general wall surfaces. We compared Stucco against the other materials to investigate whether we could improve disinfection for the shadow area. The reflectance ratios of UV-C irradiation of each material were examined, with particular attention to the rates for the author's Modified Stucco. To evaluate the disinfection effects of the UV-C reflective lighting, colonies of E. coli and of Staphylococcus hominis were cultured in an agar media and counted over a certain time period after applying UV-C irradiation from a sterilizing lamp onto the investigation materials. The author's Modified Stucco, produced reflectance ratios that was 11 times that of white wallpaper. This demonstrated that the UV-C reflected on the Stucco wall having optimum components and their compositions inhibited the number of E. coli and S. hominis, resulting in significantly disinfection effects on white wallpapers. The space with Modified Stucco and then irradiated by a UV-C may give a strong disinfection effect.

A Phase II, Multicenter, Randomized, Placebo-Controlled Study of Benralizumab, a Humanized Anti-IL-5R Alpha Monoclonal Antibody, in Patients With Eosinophilic Chronic Rhinosinusitis.

BACKGROUND: Strong eosinophil infiltration in chronic rhinosinusitis with nasal polyp (CRSwNP) is highly associated with recalcitrance and higher nasal polyp recurrence rate after surgery. The prevalence of eosinophilic CRSwNP (ECRS) is increasing in Asian countries including Japan. Benralizumab is a humanized anti-IL-5R alpha monoclonal antibody that depletes eosinophils by antibody-dependent cell-mediated cytotoxicity. OBJECTIVE: To assess the efficacy and safety of benralizumab in patients with ECRS. METHODS: This phase II, randomized, double-blind, placebo-controlled study was conducted in Japan. Patients were randomized 1:2:2 to placebo, a single administration of benralizumab 30 mg, or benralizumab 30 mg every 4 weeks (q4w) for a total of three doses. The primary endpoint was the change in nasal polyp score from baseline at Week 12. RESULTS: Overall, 56 patients were enrolled (placebo, n = 11; benralizumab single dose, n = 22; benralizumab q4w, n = 23). Although the mean total nasal polyp score began to decrease after the initiation of benralizumab treatment, there were no statistically significant differences in change in nasal polyp score from baseline at Week 12 between benralizumab and placebo (placebo, -0.5 ± 0.8; benralizumab single, -0.3 ± 0.8; benralizumab q4w, -0.5 ± 1.5). Post-hoc analysis showed that the administration of benralizumab decreased nasal polyp scores ≥2 points in 42.2% of ECRS patients and that patients with high blood eosinophil levels had a greater tendency to respond to benralizumab treatment. The safety profile was similar to that in previous studies and no unexpected adverse events were noted. CONCLUSION: Although benralizumab did not meet the primary efficacy endpoint, reductions of nasal polyp scores were seen in the benralizumab group compared with the placebo group over the whole study period, especially in patients with high levels of blood eosinophils.

Prelacrimal approach vs conventional surgery for inverted papilloma in the maxillary sinus.

BACKGROUND: The prelacrimal approach, termed endoscopic modified medial maxillectomy (EMMM), has recently been applied for treatment of inverted papilloma (IP) in the maxillary sinus. EMMM provides wider access to the maxillary sinus while preserving the inferior turbinate and nasolacrimal duct. METHODS: We reviewed patients with IP in the maxillary sinus to compare the surgical results obtained by conventional surgery (ie, endoscopic maxillary sinus antrostomy or in combination with the Caldwell-Luc approach) with those obtained by EMMM. RESULTS: All patients had a T3 on the Krouse staging system, and the average follow-up time was 46.0 months. Of the 18 patients in the conventional group, recurrence was seen in 3 patients (16.6%). No recurrence was seen in the 27 patients who showed preservation of the inferior turbinate and nasolacrimal duct, and no complications occurred in the EMMM group. CONCLUSIONS: EMMM is an effective surgical approach that reduces recurrence with fewer complications.

JESREC score and mucosal eosinophilia can predict endotypes of chronic rhinosinusitis with nasal polyps.

OBJECTIVE: Recently, JESREC score and mucosal eosinophil count have been used to diagnose eosinophilic chronic rhinosinusitis (ECRS) in Japan. However, it remains unknown whether the subtypes of CRS diagnosed by these criteria have different endotypes. In the present study, we investigated whether JESREC score and mucosal eosinophil count were appropriate for classification of CRS subgroups into endotypes. METHODS: A cross-sectional study involving 71 consecutive patients with CRS with nasal polyps (CRSwNP) and 13 control patients was performed. Nasal polyp tissues from CRSwNP patients and uncinate process tissues from control patients were collected for analysis of inflammatory cells by immunohistochemistry and measurement of cytokines and chemokines by ELISA and quantitative real-time PCR. We compared the differences between subtypes according to JESREC score and mucosal eosinophil count and investigated the subgroups with different endotypes by cluster analysis and principal component analysis. RESULTS: In the 71 CRSwNP patients, 9 patients had JESREC score <11 and mucosal eosinophil count <70/HPF (Group A), 20 patients had JESREC score ≥11 and mucosal eosinophil count <70/HPF (Group C), and 42 patients had JESREC score ≥11 and mucosal eosinophil count ≥70/high-power field (HPF) (Group D). Semiquantitative analysis of inflammatory cells showed that eosinophils, neutrophils, macrophages, mast cells, and basophils differed significantly between the subgroups. At the mRNA level, CLC, IL5, IL13, CCL11, CCL24, CCL26, POSTN, CSF3, and IL8 showed significant differences. At the protein level, eotaxin-2/CCL24, eotaxin-3/CCL26, and G-CSF had significant differences. Cluster analysis using gene expression levels in 55 CRS patients and 11 control patients revealed that the patients could be classified into five clusters. Cluster 1 (n=27) contained all patients with Group D. Cluster 2 (n=11) comprised all control patients. Cluster 3 (n=4) included mixed subtypes: one with Group A and three with Group D. Cluster 4 (n=7) and Cluster 5 (n=17) contained all patients with Groups A and C, respectively. Furthermore, the principal component analysis revealed that the subtypes had different characteristics. CONCLUSION: CRS subtypes based on JESREC score and mucosal eosinophil count showed different inflammatory patterns, and unsupervised statistical analyses supported the classification that can predict endotypes. From these results, we concluded that the classification based on JESREC score and mucosal eosinophil count was useful for predicting CRS endotypes.